Using ex vivo organotypic cultures to investigate donor cell integration in the gut

Why did we fund this project?

This award aims to use an ex vivo organotypic culture method to investigate therapeutic interventions used to treat enteric nervous system defects, reducing the number of mice in this research.

The enteric nervous system coordinates the function of the bowel including movement, hormone secretion and fluid exchange. Enteric neuropathies are degenerative conditions that can occur when the neurons making up this system degrade, and treatments are limited to managing symptoms. It has previously been shown that lost or damaged enteric nerve cells can be rescued in mice by transplanting gut nervous stem cells. However, the underlying mechanisms of how the stem cells integrate is not well understood. Researching stem cell treatment and integration typically requires the stem cells to be implanted using surgical procedures in vivo, usually using mice. Dr Conor McCann and colleagues have developed a culture method that allows portions of the murine gut to be grown ex vivo for up to three weeks. Importantly, six portions of bowel can be obtained from one mouse gut, reducing the number of mice needed overall for this type of research.

With NC3Rs funding, Conor will build confidence in the ex vivo culture method and demonstrate its utility in cell-based transplantation applications. Mouse stem cells will be tagged with a fluorescent marker and introduced into the ex vivo model to track the stem cells after implantation. Conor will then determine if these stem cells integrate into the ex vivo tissue. The transcriptomics of the gut before and after implantation will be analysed before the functionality of the implanted stem cells is determined.